Norepinephrine-induced beta 1-adrenergic peripheral vasodilation in conscious dogs.
Norepinephrine (NE) elicits alpha-adrenergic vasoconstriction and beta 1-adrenergic increases in heart rate and myocardial contractility. To determine whether NE can also elicit peripheral beta 1-adrenergic vasodilation, conscious dogs were studied after recovery from instrumentation for the measurement of cardiac output, arterial pressure, and left ventricular (LV) pressure and calculations of LV dP/dt and total peripheral resistance (TPR). NE, after pretreatment with hexamethonium and phentolamine, reduced mean arterial pressure 40 +/- 5% from 117 +/- 9 mmHg and TPR 60 +/- 5% from 0.058 +/- 0.007 mmHg X ml-1 X min and increased cardiac output 55 +/- 11% from 2,159 +/- 188 ml/min. beta 1-Adrenergic blockade with atenolol reversed the vasodilation induced by NE completely, while at this time isoproterenol was still able to reduce peripheral resistance further, by 67 +/- 7%, indicating that beta 2-adrenergic receptors were not blocked. Administration of phentolamine to intact dogs caused a fall in mean arterial pressure (23 +/- 5%) and TPR (34 +/- 5.4%) and an endogenous increase in plasma NE (2,987 +/- 905 pg/ml) and epinephrine (584 +/- 92 pg/ml). These increases in cardiac output and decreases in TPR were also reversed by atenolol (0.5 mg/kg). Moreover, this dose of atenolol blocked the increases in iliac blood flow induced by local injection of NE in the limb. Thus, in the presence of alpha-adrenergic receptor blockade, either administration of NE or release of endogenous NE elicits potent peripheral vasodilation, which appears to involve a beta 1-adrenergic receptor mechanism.


The Doctor-Patient Relationship With Artificial Intelligence.
The doctor-patient relationship has been evolving from benevolent paternalism to a more patient-centered relationship in the modern era. Although artificial intelligence (AI) has the potential to improve nearly every aspect of health care, many physicians are skeptical about integrating AI into their current medical practice. The purpose of this article is to explore what AI means for the doctor-patient relationship and for breast imaging radiologists. The promise of AI is its potential to release physicians from tasks that are better performed by automation. AI may enhance our diagnostic accuracy to the point that we are able to refocus on the art of the doctor-patient relationship.


Tunable nonuniform sampling method for fast calculation and intensity modulation in 3D dynamic holographic display.
Heavy computational load of computer-generated hologram (CGH) and imprecise intensity modulation of 3D images are crucial problems in dynamic holographic display. The nonuniform sampling method is proposed to speed up CGH generation and precisely modulate the reconstructed intensities of phase-only CGH. The proposed method can eliminate the redundant information properly, where 70% reduction in the storage amount can be reached when it is combined with the novel lookup table method. Multigrayscale modulation of reconstructed 3D images can be achieved successfully. Numerical simulations and optical experiments are performed, and both are in good agreement. It is believed that the proposed method can be used in 3D dynamic holographic display.


Can an ankle-foot orthosis change hearts and minds?
The current military conflicts of Operation Enduring Freedom and Operation Iraqi Freedom have been characterized by high-energy explosive wounding patterns, with the majority affecting the extremities. While many injuries have resulted in amputation, surgical advances have allowed the orthopaedic surgeon to pursue limb salvage in the face of injuries once considered unsalvageable. The military limb salvage patient is frequently highly active and motivated and expresses significant frustration with the slow nature of limb salvage rehabilitation and continued functional deficits. Inspired by these patients, efforts at this institution began to provide them with a more dynamic orthosis. Utilizing techniques and technology resulting from cerebral palsy, stroke, and amputation research, the Intrepid Dynamic Exoskeletal Orthosis was created. To date, this device has significantly improved the functional capabilities of the limb salvage wounded warrior population when combined with a high-intensity rehabilitation program. Clinical and biomechanical research is currently underway at this institution in order to fully characterize the device, its effect on patients, and what can be done to modify future generations of the device to best serve the combat-wounded limb salvage population.


Comparison of adjuvant therapy for node-positive clinical stage IB-IIB cervical cancer: Systemic chemotherapy versus pelvic irradiation.
This was a nation-wide retrospective study in Japan examining women who underwent radical hysterectomy for clinical stage IB-IIB cervical cancer with pelvic and/or para-aortic lymph node metastasis between 2004 and 2008. Time to recurrence or death and patterns of disease recurrence were compared based upon the adjuvant treatment pattern: whole pelvic radiotherapy alone (n = 253), concurrent chemoradiotherapy (CCRT, n = 502) and chemotherapy alone (n = 319). Women who received chemotherapy alone had similar recurrence (5-year rates, 36.6% vs. 34.1%, adjusted-hazard ratio [HR] 0.95, 95% confidence interval [CI] 0.70-1.28, P = 0.72) and cervical cancer mortality (24.7% vs. 21.8%, adjusted-HR 0.96, 95% CI 0.67-1.38, P = 0.83) rates compared to those who received CCRT on multivariate analysis. However, when recurrence patterns were stratified, chemotherapy treatment was independently associated with decreased risk of distant recurrence (5-year cumulative rates, 19.2% vs. 24.6%, adjusted-HR 0.47, 95% CI 0.31-0.71, P < 0.001) but increased risk of local recurrence (23.9% vs. 14.3%, adjusted-HR 2.03, 95% CI 1.34-3.08, P = 0.001) compared to CCRT. Non-squamous histology, parametrial involvement and high lymph node ratio were independent predictors for local recurrence, and presence of multiple risk factors was associated with high 5-year cumulative local recurrence rate in the chemotherapy group: no risk factor 3.9%, single factor 14.2-22.1%, and multiple risk factors 27.8-71.9% (P < 0.001). In conclusion, while exhibiting different recurrence patterns, systemic chemotherapy may be as effective a postoperative treatment as radiation-based therapy in node-positive high-risk stage IB-IIB cervical cancer. When tumor exhibits certain risk factors, chemotherapy alone is likely insufficient for local control and adding pelvic irradiation to systemic chemotherapy is recommended in this subgroup.


Analysis and prediction of leucine-rich nuclear export signals.
We present a thorough analysis of nuclear export signals and a prediction server, which we have made publicly available. The machine learning prediction method is a significant improvement over the generally used consensus patterns. Nuclear export signals (NESs) are extremely important regulators of the subcellular location of proteins. This regulation has an impact on transcription and other nuclear processes, which are fundamental to the viability of the cell. NESs are studied in relation to cancer, the cell cycle, cell differentiation and other important aspects of molecular biology. Our conclusion from this analysis is that the most important properties of NESs are accessibility and flexibility allowing relevant proteins to interact with the signal. Furthermore, we show that not only the known hydrophobic residues are important in defining a nuclear export signals. We employ both neural networks and hidden Markov models in the prediction algorithm and verify the method on the most recently discovered NESs. The NES predictor (NetNES) is made available for general use at http://www.cbs.dtu.dk/.


tPA promotes cortical neuron survival via mTOR-dependent mechanisms.
Tissue plasminogen activator (tPA) is a thrombolytic agent commonly used in the treatment of ischemic stroke. While the thrombolytic effects of tPA have been well established, the impact of this blood-brain barrier (BBB) crossing drug on neurons is not known. Given the widespread use of tPA in the clinical setting and the strict therapeutic window established for effective use of the drug, we examined the molecular mechanisms mediating the impact of tPA on postnatal cortical neurons isolated from the mouse brain. Dissociated postnatal primary cortical neurons were treated with tPA and the effects on neuron survival were evaluated. Pharmacological inhibitors of several signaling pathways previously implicated in neuroprotection (mTOR, JAK/STAT, MAPK and PKA-dependent mechanisms) were used to pinpoint the mechanistic effectors of tPA on neuron survival in vitro. We report here that tPA treatment results in a time-dependent neuroprotective effect on postnatal cortical neurons that relies predominantly on Janus kinase (JAK) and mammalian target of rapamycin (mTOR) signaling mechanisms. Taken together, these data suggest that tPA promotes neuroprotection in a temporally-regulated manner and that both JAK and mTOR signaling effectors are critical mediators of this neuroprotective effect. The results suggest the possibility of targeting these defined mechanisms to potentially expand the therapeutic window for tPA.


What Factors Are Associated with Increased Financial Burden and High Financial Worry For Patients Undergoing Common Hand Procedures?
Few studies have examined whether orthopaedic surgery, including hand surgery, is associated with patients' financial health. We sought to understand the level of financial burden and worry for patients undergoing two common hand procedures-carpal tunnel release and open reduction and internal fixation for a distal radius fracture-as well as to determine factors associated with a higher financial burden and worry. In patients undergoing operative treatment for isolated carpal tunnel syndrome with carpal tunnel release or open reduction and internal fixation for a distal radius fracture, we used validated financial burden and worry questionnaires to ask: (1) What percentage of patients report some level of financial burden, and what is the median financial burden composite score? (2) What percentage of patients report some level of financial worry, and what percentage of patients report a high level of financial worry? (3) When accounting for other assessed factors, what patient- and condition-related factors are associated with financial burden? (4) When accounting for other assessed factors, what patient- and condition-related factors are associated with high financial worry? In this cross-sectional survey study, a hand and upper extremity database at a single tertiary academic medical center was reviewed for patients 18 years or older undergoing operative treatment in our hand and upper extremity division for an isolated distal radius fracture between October 2017 and October 2019. We then selected all patients undergoing carpal tunnel release during the first half of that time period (given the frequency of carpal tunnel syndrome, a 1-year period was sufficient to ensure comparable patient groups). A total of 645 patients were identified (carpal tunnel release: 60% [384 of 645 patients]; open reduction and internal fixation for a distal radius fracture: 40% [261 of 645 patients). Of the patients who underwent carpal tunnel release, 6% (24 of 384) were excluded because of associated injuries. Of the patients undergoing open reduction and internal fixation for a distal radius fracture, 4% (10 of 261) were excluded because of associated injuries. All remaining 611 patients were approached. Thirty-six percent (223 of 611; carpal tunnel release: 36% [128 of 360]; open reduction and internal fixation: 38% [95 of 251]) of patients ultimately completed two validated financial health surveys: the financial burden composite and financial worry questionnaires. Descriptive statistics were calculated to report the percentage of patients who had some level of financial burden and worry. Further, the median financial burden composite score was determined. The percentage of patients who reported a high level of financial worry was calculated. A forward stepwise regression model approach was used; thus, variables with p values < 0.10 in bivariate analysis were included in the final regression analyses to determine which patient- and condition-related factors were associated with financial burden or high financial worry, accounting for all other measured variables. The median financial burden composite score was 0 (range 0 [lowest possible financial burden] to 6 [highest possible financial burden]), and 13% of patients (30 of 223) reported a high level of financial worry. After controlling for potentially confounding variables like age, insurance type, and self-reported race, the number of dependents (regression coefficient 0.15 [95% CI 0.008 to 0.29]; p = 0.04) was associated with higher levels of financial burden, while retired employment status (regression coefficient -1.24 [95% CI -1.88 to -0.60]; p < 0.001) was associated with lower levels of financial burden. In addition, the number of dependents (odds ratio 1.77 [95% CI 1.21 to 2.61]; p = 0.004) and unable to work or disabled employment status (OR 3.76 [95% CI 1.25 to 11.28]; p = 0.02) were associated with increased odds of high financial worry. A notable number of patients undergoing operative hand care for two common conditions reported some degree of financial burden and worry. Patients at higher risk of financial burden and/or worry may benefit from increased resources during their hand care journey, including social work consultation and financial counselors. This is especially true given the association between number of dependents and work status on financial burden and high financial worry. However, future research is needed to determine the return on investment of this resource utilization on patient clinical outcomes, overall quality of life, and well-being. Level III, therapeutic study.


Allicin has significant effect on autoimmune anti-islet cell antibodies in type 1 diabetic rats.
The research purpose was to experimentally investigate the effect of allicin administration on the levels of main type 1 diabetes (IDDM) autoantibodies which are anti-islet cell antibodies (ICA) with an attempt to find a relation between this immunological effect and histological and/or biochemical findings. We have evaluated, with the help of ELISA kits, the levels of ICA and serum insulin in male Sprague-Dawley rats with Streptozotocin-induced IDDM in addition to pancreatic histological findings. The four groups (6 rats each) under study received or not different intraperitoneal doses of allicin for a period of 30 days. Daily intraperitoneal administration of allicin (either at as low dose of 8 mg/kg or high dose of 16 mg/kg) for up to 30 days to type 1 diabetic rats effectively reduces levels of anti-islet cell antibodies and in addition, reduced the level of insulin due to damaged Langerhans islet cell was significantly increased in the serum due to a repairing tissue process in pancreatic tissues. These experimental results suggest that allicin treatment has a therapeutic protective effect against autoimmune reactions occurring in IDDM. The data may provide new strategies for using allicin to be recommended as an excellent candidate in the clinical management, control, and prevention of IDDM.


Recurrent oil sheens at the deepwater horizon disaster site fingerprinted with synthetic hydrocarbon drilling fluids.
We used alkenes commonly found in synthetic drilling-fluids to identify sources of oil sheens that were first observed in September 2012 close to the Deepwater Horizon (DWH) disaster site, more than two years after the Macondo well (MW) was sealed. While explorations of the sea floor by BP confirmed that the well was sound, they identified the likely source as leakage from an 80-ton cofferdam, abandoned during the operation to control the MW in May 2010. We acquired sheen samples and cofferdam oil and analyzed them using comprehensive two-dimensional gas chromatography. This allowed for the identification of drilling-fluid C16- to C18-alkenes in sheen samples that were absent in cofferdam oil. Furthermore, the spatial pattern of evaporative losses of sheen oil alkanes indicated that oil surfaced closer to the DWH wreckage than the cofferdam site. Last, ratios of alkenes and oil hydrocarbons pointed to a common source of oil found in sheen samples and recovered from oil-covered DWH debris collected shortly after the explosion. These lines of evidence suggest that the observed sheens do not originate from the MW, cofferdam, or from natural seeps. Rather, the likely source is oil in tanks and pits on the DWH wreckage, representing a finite oil volume for leakage.


DNA adduct formation and oxidative stress in colon and liver of Big Blue rats after dietary exposure to diesel particles.
Exposure to diesel exhaust particles (DEP) via the gastrointestinal route may impose risk of cancer in the colon and liver. We investigated the effects of DEP given in the diet to Big Blue rats by quantifying a panel of markers of DNA damage and repair, mutation, oxidative damage to proteins and lipids, and antioxidative defence mechanisms in colon mucosa cells, liver tissue and the blood compartment. Seven groups of rats were fed a diet with 0, 0.2, 0.8, 2, 8, 20 or 80 mg DEP/kg feed for 21 days. DEP induced a significant increase in DNA strand breaks in colon and liver. There was no effect on oxidative DNA damage (8-oxodG) in colon or liver DNA or in the urine. However, the mRNA expression of OGG1, encoding an enzyme involved in repair of 8-oxodG, was increased by DEP in both liver and colon. DNA adduct levels measured by 32P-post-labelling were elevated in colon and liver, and the expression of ERCC1 gene was affected in liver, but not in colon. In addition to these effects, DEP exposure induced apoptosis in liver. There was no significant change in mutation frequency in colon or liver. The levels of oxidative protein modifications (oxidized arginine and proline residues) were increased in liver accompanied by enhanced vitamin C levels. In plasma, we found no significant effects on oxidative damage to proteins and lipids, antioxidant enzymes or vitamin C levels. Our data indicate that gastrointestinal exposure to DEP induces DNA adducts and oxidative stress resulting in DNA strand breaks, enhanced repair capacity of oxidative base damage, apoptosis and protein oxidation in colon mucosa cells and liver.


Cluster algorithms for frustrated two-dimensional Ising antiferromagnets via dual worm constructions.
We report on the development of two dual worm constructions that lead to cluster algorithms for efficient and ergodic Monte Carlo simulations of frustrated Ising models with arbitrary two-spin interactions that extend up to third-neighbors on the triangular lattice. One of these algorithms generalizes readily to other frustrated systems, such as Ising antiferromagnets on the Kagome lattice with further neighbor couplings. We characterize the performance of both these algorithms in a challenging regime with power-law correlations at finite wave vector.


Reactive oxygen species as mediators of sperm capacitation and pathological damage.
Oxidative stress plays a major role in the life and death of mammalian spermatozoa. These gametes are professional generators of reactive oxygen species (ROS), which appear to derive from three potential sources: sperm mitochondria, cytosolic L-amino acid oxidases, and plasma membrane Nicotinamide adenine dinucleotide phosphate oxidases. The oxidative stress created via these sources appears to play a significant role in driving the physiological changes associated with sperm capacitation through the stimulation of a cyclic adenosine monophosphate/Protein kinase A phosphorylation cascade, including the activation of Extracellular signal regulated kinase-like proteins, massive up-regulation of tyrosine phosphorylation in the sperm tail, as well as the induction of sterol oxidation. When generated in excess, however, ROS can induce lipid peroxidation that, in turn, disrupts membrane characteristics that are critical for the maintenance of sperm function, including the capacity to fertilize an egg. Furthermore, the lipid aldehydes generated as a consequence of lipid peroxidation bind to proteins in the mitochondrial electron transport chain, triggering yet more ROS generation in a self-perpetuating cycle. The high levels of oxidative stress created as a result of this process ultimately damage the DNA in the sperm nucleus; indeed, DNA damage in the male germ line appears to be predominantly induced oxidatively, reflecting the vulnerability of these cells to such stress. Extensive evaluation of antioxidants that protect the spermatozoa against oxidative stress while permitting the normal reduction-oxidation regulation of sperm capacitation is therefore currently being undertaken, and has already proven efficacious in animal models.


Similarities and differences between renal sympathetic denervation and carotid baroreceptor stimulation.
The two novel approaches recently introduced for the treatment of resistant hypertension, i.e. carotid baroreceptor stimulation and renal denervation, share a number of similarities but are also characterized by important differences. The similarities include the evidence that both interventions have as common pathophysiological background the state of sympathetic overdrive characterizing essential hypertension. In addition both procedures 1) are invasive, 2) exert in the short-term period clearcut blood pressure lowering effects and 3) still face a number of open questions, particularly related to the long-term blood pressure lowering effects, impact on end-organ damage and on cardiovascular events. The differences include the fact that two procedures act on distinct targets that trigger sympathetic activation and consequently blood pressure increase. In addition, only in the case of carotid baroreceptor stimulation the blood pressure effects can be easily assessed immediately following the implantation. Finally, the economic costs, metabolic effects and impact on vagal modulation of heart rate are different between the two interventions. This paper will provide a comparison of the background, effects and outcome of renal denervation and carotid baroreceptor stimulation, stressing whenever possible the clinical implications of the main features of the two interventions.


Isolated splenic vein thrombosis.
Isolated splenic vein thrombosis may lead to a specific clinical presentation, namely, bleeding from isolated gastric varices (which are difficult to diagnose), splenomegaly, and normal liver function. The most common cause is pancreatic disease. To exclude associated portal venous thrombosis, angiography is essential prior to a splenectomy, which is curative of the variceal hemorrhage. Splenic vein thrombosis may also be noted as an incidental finding during angiography. Although the natural history is uncertain, a splenectomy is probably not indicated until a hemorrhage occurs.


A new variant of sensory ataxic neuropathy with autosomal dominant inheritance.
We describe a Dutch family with sensory ataxia in two generations, late onset of symptoms (over the age of 40 years) and slow progression. Clinical, electrophysiological and sural nerve biopsy findings revealed a sensory polyneuropathy due to axonal degeneration of myelinated nerve fibres in four of five investigated siblings. Other neurological abnormalities in the affected family members consisted only of mild eye movement disturbances, probably due to cerebellar involvement. Five other family members were investigated and found unaffected. As the disease is inherited from the affected father to his sons and daughters, this is the first description of a probably autosomal dominant form of late onset hereditary sensory neuropathy with predominant sensory ataxia and minor other neurological abnormalities.


Management of labour and delivery in congenitally corrected transposition of great arteries.
A descriptive case report of the labour and delivery management of a 28-year-old woman who presented with congenitally corrected transposition of great arteries, dextrocardia, systemic ventricular dysfunction and junctional tachycardia. Patients with congenitally corrected transposition have a thin-walled morphological right ventricle as the systemic circulatory pump. The stress of increased cardiac output can lead to congestive heart failure, systemic atrioventricular valve regurgitation and arrhythmias. We used minimally invasive continuous cardiac output monitoring, fluid balance optimization and good maternal pain control to prevent decompensation and achieve vaginal delivery with a good maternal and neonatal outcome.


[Evaluating the effectiveness of a fixed combination of amlodipine and bisoprolol in ambulatory patients with arterial hypertension and ischemic heart disease].
To evaluate the efficacy and safety of a fixed-term combination of β-blocker bisoprolol and the calcium antagonist amlodipine in the outpatient treatment of patients with arterial hypertension (AH) and coronary heart disease (CHD). The study involved 100 patients with hypertension 1-3 first degree and CHD, which previous antihypertensive therapy was ineffective or irregular. Inclusion in the treatment regimen of amlodipine was associated with significant decrease in blood pressure (BP) with the achievement of the target level of systolic blood pressure of 90%, diastolic blood pressure--in 97% of cases after 4 weeks treatment. Analysis of the dynamics of the heart rate during the visits to the doctor and the patient diary data revealed a significant slowing on the background of the therapy, which is important for patients with coronary artery disease. In carrying out daily monitoring of ECG Holter dynamics observed a decrease in the average duration of episodes of depression segment ST on 50.5% (p < 0.05) and the number of ischemic episodes at 54.8% (p < 0.05). The use of fixed-dose combination of bisoprolol and amlodipine allowed to increase the compliance of patients with treatment 2.2 times.


Corticosteroids in the emergency department therapy of acute adult asthma: an evidence-based evaluation.
To review the literature to determine the benefits of corticosteroids (CCSs) (oral, IM, IV, or inhaled) in the treatment of adult patients with acute asthma presenting at an acute-care setting. A MEDLINE search was conducted using the following terms: (1) Asthma OR Wheez, AND (2) Glucocorticoids OR Steroids, AND (3) Acute OR Emerg. Other sources were the CURRENT CONTENTS database, review articles, reference sections of located studies, and a manual search of the top 15 journals for respiratory and emergency medicine. Patients were selected for the study by the following criteria: (1) English language; (2) adult patients with asthma whose acute exacerbations were the primary reason for assessment; (3) involvement in randomized, controlled trials conducted in an emergency care setting; (4) patients had participated in a study investigating a primary research question involving treatment with CCSs; and (5) outcomes based on results of pulmonary function tests and on hospital admission rates. At the 3-h assessment, only high doses of inhaled CCSs significantly improved pulmonary function compared with placebo (effect size [ES], 0.56; 95% confidence interval [CI], 0.15 to 0.97). On the other hand, after receiving IV CCSs, patients required at least 6 to 24 h to show moderate but nonsignificant improvements of pulmonary function (6-h ES, 0.44 [95% CI, -0.01 to 0.89]; 12-h ES, 0.54 [95% CI, -0.08 to 1.17]; and 24-h ES, 0.53 [95% CI, -0.39 to 1.45]). The data from the six studies that we used to pool information on admission rate outcome showed a 32% reduction in favor of the use of IV CCSs (relative risk [RR], 0.68 [95% CI, 0.47 to 0.99]; number needed to treat, 12.5 [95% CI, 7.1 to 50]). However, the pooled effect of the three high-quality studies showed no difference between groups (RR, 1.21; 95% CI, 0.67 to 2.18). Oral CCSs provided a similarly beneficial effect on pulmonary function when compared with parenteral administration (ES, -0.14; 95% CI, -0.82 to 0.31. Finally, the results showed a nonsignificant favorable trend toward improved outcome with medium or high doses of CCSs. This evidence-based evaluation suggests that the administration of parenteral CCSs to the patient on arrival at the emergency department (ED) neither improves airflow obstruction nor reduces the need for hospitalization. Parenteral CCSs probably require >6 to 24 h to begin to act. Comprehensible conclusions about admission rates in the ED setting are difficult to make. At the 3-h assessment, only high doses of inhaled CCSs (in one study) significantly improved pulmonary function compared with placebo. IV and oral CCSs appear to have equivalent effects, and there is a tendency toward improvement in pulmonary function with medium or high doses.


Emerging therapeutic targets for idiopathic pulmonary fibrosis: preclinical progress and therapeutic implications.
Idiopathic pulmonary fibrosis (IPF) is a chronic progressive lung disease with high associated morbidity and mortality. The therapeutic landscape has significantly changed in the last 20 years with two drugs currently approved that have demonstrated the ability to slow disease progression. Despite these developments, survival in IPF is limited, so there is a major interest in therapeutic targets which could serve to open up new therapeutic avenues. We review the most recent information regarding drug targets and therapies currently being investigated in preclinical and early-stage clinical trials. The complex pathogenesis of IPF and variability in disease course and response to therapy highlights the importance of a precision approach to therapy. Novel technologies including transcriptomics and the use of serum biomarkers, will become essential tools to guide future drug development and therapeutic decision making particularly as it pertains to combination therapy.


Effects of ADP-ribosylation of GTP-binding protein by pertussis toxin on immunoglobulin E-dependent and -independent histamine release from mast cells and basophils.
Pretreatment of rat peritoneal mast cells, human basophils, bone marrow-derived mouse mast cells (BMMC) and mouse mast cell line PT-18 cells with 1 microgram/ml pertussis toxin (PT) failed to inhibit immunoglobulin E (IgE)-dependent histamine release from the cells. In BMMC and PT-18 cells, even 20-hr incubation of the cells with 1 microgram/ml PT, which ADP-ribosylates more than 97% of 41 kDa, alpha-subunit of Ni in the cells, failed to affect the IgE-dependent release of histamine or arachidonate. The results indicate that GTP-binding protein, Ni, is not involved in the transduction of triggering signals induced by cross-linking of IgE receptors. In contrast, pretreatment of rat mast cells with 1 ng/ml to 0.1 microgram/ml PT for 2 hr inhibited histamine release induced by compound 48/80 in a dose-dependent manner. A similar pretreatment with PT inhibited thrombin-induced histamine release from BMMC and N-formyl-L-methionyl-L-leucyl-L-phenylalanine-induced histamine release from human basophils in a similar dose-dependent fashion. However, even 20 hr of incubation of sensitized BMMC with 1 microgram/ml PT failed to inhibit either thrombin-induced or antigen-induced breakdown of phosphatidylinositides (PI), i.e., the formation of inositol triphosphate and diacylglycerol, Quin-2 signal, and the release of arachidonic acid. The results indicate that the inhibition of thrombin-induced histamine release by PT-treatment is not due to the inhibition of PI-turnover, and that Ni is not involved in thrombin-induced or antigen-induced (IgE-dependent) hydrolysis of phosphatidylinositides in mast cells.


Biological dosimetry for triage of casualties in a large-scale radiological emergency:capacity of the EU member states.
In recent years, a number of events have occurred that highlight the necessity of being prepared for a possible large-scale radiological event. An important question is how well are European Union (EU) Member States prepared to cope with mass radiological casualties. A survey to establish the current status of biological dosimetry across the EU was carried out with the aim of assessing capacity to perform biodosimetric triage of accident victims. Information was sought from the radiation protection authorities of the 27 Member States plus Switzerland and Norway. Biology dosimetry is established in 15 EU countries. Their total capacity for dosimetric triage is about 1500 cases per week analysed with a dicentric assay or about 800 cases analysed with a micronucleus assay. Although these numbers appear encouraging, there is not much collaboration between the laboratories and what is required is a network at the EU level.


[Use of mathematical models in the analysis of gastric digestion].
Mathematical models of gastric hydrolysis of different proteins were elaborated in dogs. These models consider the spatialtemporal coordinates of distribution of pepsin, hydrogen ions and hydrolysates in the stomach, carry out the analysis of factors determining the gastric digestion. The main part belongs to topography and hydrodynamics of gastric content as well as to pepsin concentration. The concentration of "total hydrolysates" is proposed as the final criterion of the gastric digestion. An algorithm for mathematical modelling of gastric digestion is suggested.


Analysis of proteins from different phase variants of the entomopathogenic bacteria Photorhabdus luminescens by two-dimensional zymography.
Two-dimensional zymography which combines two-dimensional electrophoresis with zymography was used to analyze proteases and other proteins produced by different phase variants of two strains of Photorhabdus luminescens. Both the primary and secondary phases of P. luminescens strains Hp and Hm secreted proteases. The protease in P. luminescens Hp has a molecular weight (Mr) of 57,000 and an isoelectric point (pI) of 4.4 whereas that in P. luminescens Hm has an Mr of 59,000 and pI of 4.9. Several putative protease degradation products were clearly visible in the zymograms from both bacterial strains. Two-dimensional zymography also showed that several secretory proteins were present only in particular phase variants and therefore could be used as specific markers. Unexpectedly, the two-dimensional zymography revealed that a nonsecretory protease with an Mr of 47,000 and a pI of 4.0 was present in the cell extracts of all phases of both P. luminescens Hp and Hm. The application of the two-dimensional zymography for the identification of other enzymes was also discussed.


SATB1 Knockdown Inhibits Proliferation and Invasion and Decreases Chemoradiation Resistance in Nasopharyngeal Carcinoma Cells by Reversing EMT and Suppressing MMP-9.
Background: Special AT-rich sequence binding protein 1 (SATB1) is a chromatin organizer and transcriptional regulator which regulate numerous cellular processes through effects on multiple gene expression. SATB1 is associated with drug resistance in several cancers. Whether SATB1 involves radiation resistance in nasopharyngeal carcinoma (NPC) and underlying mechanism of SATB1 to participate in chemoradiotherapy resistance in NPC have not been elaborated. Methods: Chemoradioresistant NPC cell lines 5-8F/DDP (cisplatin) and 5-8F/R (radiation) were developed from 5-8F cell line. The expressions of SATB1, MMP-9 and EMT markers (Vimentin and E-cadherin) in these cell lines were examined by reverse transcription-quantitative (RT-q) PCR and western blot (WB) analysis. Cell viabilities of 5-8F/DDP treated with various concentrations of DDP and 5-8F/R irradiated with various doses of X-ray at the indicated time were investigated by MTT test. SATB1 was silenced in 5-8F/DDP and 5-8F/R cells by short hairpin RNA, and then the expressions of SATB1, MMP-9, Vimentin and E-cadherin were evaluated by RT-qPCR and WB analysis; the abilities of cell proliferation and invasion were assessed using MTT and transwell assays, respectively. Drug and radiation resistance assays were performed after SATB1 knockdown and cell viability was detected by MTT method. Results: SATB1, MMP-9 and Vimentin were markedly upregulated in 5-8F/DDP and 5-8F/R cells compared with 5-8F cell, whereas E-cadherin was obviously downregulated. 5-8F/DDP and 5-8F/R cells displayed drug and radiation resistance to DDP or X-irradiation, respectively, while DDP or X-irradiation inhibited 5-8F cell viability in a time- and dose-dependent manner. Subsequently, knockdown of SATB1 resulted in decreased MMP-9 and Vimentin expression and increased E-cadherin expression in 5-8F/DDP and 5-8F/R. Furthermore, silencing of SATB1 suppressed proliferative and invasive abilities of 5-8F/DDP and 5-8F/R cells. Additionally, SATB1 knockdown reduced drug resistance of 5-8F/DDP cell to DDP and decreased radiation resistance of 5-8F/R cell to X-ray. Conclusion: These results suggest that high expression of SATB1 plays an important role in the malignant behavior of NPC and leads to X-radiation and drug resistance in NPC through promoting EMT process and enhancing MMP-9 expression. SATB1 may be a promising therapeutic target for aggressive and chemoradiation resistant NPC.


Comparison of radiographic parameters for analysis of normal and dysplastic hips in the adult.
Radiographic quantitation of the dysplastic hip in adults is difficult. This study compares the values for commonly used indices, the acetabular angle (AA), the center edge angle (CEA), and femoral head coverage, and the X-Y coordinate system on the anteroposterior pelvic radiograph in 30 adult patients with 60 normal hips, and 20 adult patients with 27 dysplastic hips. Dysplastic hips demonstrated significantly higher values for the AA and the X-Y coordinates, and significantly lower values for the CEA and femoral head coverage compared to normal hips. Femoral head coverage in dysplastic hips correlated best (negatively) with the Y coordinate, i.e., poorer coverage was associated with greater superior migration of the femoral head. The AA describes the slope of the acetabular roof, but does not take into account the relative position of the femoral head. The CEA measures the position of the femoral head in relation to the lateral lip of the acetabulum, but does not necessarily use the true acetabulum. The X-Y coordinate system relates the center of the femoral head to an identifiable acetabular landmark, the teardrop shadow. These coordinates are easily determined and can serve as an adjunct to other radiographic indices that quantitate the amount of subluxation in dysplastic hips in adults.


Multi-omic dissection of the drought resistance traits of soybean landrace LX.
With diverse genetic backgrounds, soybean landraces are valuable resource for breeding programs. Herein, we apply multi-omic approaches to extensively characterize the molecular basis of drought tolerance in the soybean landrace LX. Initial screens established that LX performed better with PEG6000 treatment than control cultivars. LX germinated better than William 82 under drought conditions and accumulated more anthocyanin and flavonoids. Untargeted mass spectrometry in combination with transcriptomic analyses revealed the chemical diversity and genetic basis underlying the overall performance of LX landrace. Under control and drought conditions, significant differences in the expression of a suite of secondary metabolism genes, particularly those involved in the general phenylpropanoid pathway and flavonoid but not lignin biosynthesis, were seen in LX and William 82. The expression of these genes correlated with the corresponding metabolites in LX plants. Further correlation analysis between metabolites and transcripts identified pathway structural genes and transcription factors likely are responsible for the LX agronomic traits. The activities of some key biosynthetic genes or regulators were confirmed through heterologous expression in transgenic Arabidopsis and hairy root transformation in soybean. We propose a regulatory mechanism based on flavonoid secondary metabolism and adaptive traits of this landrace which could be of relevance to cultivated soybean.


The structure of a DNA unwinding protein and its complexes with oligodeoxynucleotides by x-ray diffraction.
The structure of the gene 5 DNA unwinding protein from bacteriophage fd has been solved to 2.3 A resolution by x-ray diffraction techniques. The molecule contains an extensive cleft region that we have identified as the DNA binding site on the basis of the residues that comprise its surface. The interior of the groove has a rather large number of basic amino acid residues that serve to draw the polynucleotide backbone into the cleft. Arrayed along the external edges of the groove are a number of aromatic amino acid side groups that are in position to stack upon the bases of the DNA and fix it in place. The cleft then acts as an elongated pair of jaws that draws the DNA between them by charge interactions involving the phosphates with the interior lysines and arginines. The jaws then close on the DNA strand through small conformation changes and the rotation of aromatic side-chains into position to stack upon the purines and pyrimidines. Complexes of the gene 5 protein with a variety of oligodeoxynucleotides have been formed and crystallized for x-ray diffraction analysis. The crystallographic parameters of four different unit cells indicate that the fundamental unit of the complex is composed of six gene 5 protein dimers. We believe this aggregate has 622 point group symmetry and is a ring formed by end to end closure of a linear array of six dimers. From our results we have proposed a double helical model for the gene 5 protein-DNA complex in which the protein forms a spindle or core around which the DNA is spooled. 5.0-A x-ray diffraction data from one of the crystalline complexes is currently being analyzed by molecular replacement techniques to obtain what we believe will be the first direct visualization of a protein-deoxyribonucleic acid complex approaching atomic resolution.


Histopathological and biochemical findings of congenital copper deficiency: are these similar to those of caprine arthritis-encephalitis?
This study was done after identifying animals with a twisted carpal joint in goat herd. These included a kid goat walking on its articulus carpii and a newborn goat with a stiff leg. Necropsies of the diseased goats revealed swollen carpal joints that were twisted backwards. Arthritis was observed during microscopic examination of the carpal joints. Very low levels of eosinophil, leucocyte, and lymphocyte cell infiltration were found in the central nervous system and meninges. Serum copper levels were significantly decreased in most of the animals. All of these results led us to diagnose the animals with swayback disease.


Application of a multi-faceted approach for the assessment of treatment response in falciparum malaria: a study from Malaysian Borneo.
Thirty-two patients reporting to the Lundu District Hospital, Sarawak, Malaysian Borneo, with uncomplicated falciparum malaria were recruited into a multifaceted study to assess treatment response. Following combined chloroquine and sulphadoxine/pyrimethamine treatment the patients were followed for 28 days according to the World Health Organisation in vivo drug response protocol. The in vivo study revealed that 13 (41%) of the patients had a sensitive response to treatment, five (16%) cleared asexual stage parasites but had persistent gametocytes, 11 (34%) had RI type resistance and three (9%) had RII type resistance requiring quinine intervention before day 7 for parasite clearance. Although clinically insignificant, patients with persistent gametocytes, surviving chloroquine and sulphadoxine/pyrimethamine treatment during maturation, were placed in the reduced response to treatment group for analysis. Allelic typing detected 100% prevalence of the pfcrt K76T marker associated with chloroquine resistance and 78% prevalence of the pfdhfr NRNL haplotype associated with sulphadoxine/pyrimethamine treatment failure. High serum chloroquine levels and pfdhfr haplotypes with <or=2 point mutations were independently associated with a sensitive response to treatment. However four patients in the sensitive response group had both low serum chloroquine levels and the pfdhfr NRNL haplotype. A degree of immunity, consistent with malaria endemicity, may account for the apparently paradoxical presence of markers of drug resistance in the sensitive group.


Use of primate model system to identify Chlamydia trachomatis protein antigens recognized uniquely in the context of infection.
A primate model system was used to identify Chlamydia trachomatis antigens uniquely recognized in the context of infection. Serum antibody titres were measured in cynomolgus monkeys challenged urethrally with C. trachomatis serovar L2 elementary bodies (EBs). High-titre sera from these primates were used, in parallel with antisera against killed C. trachomatis EBs, to differentially screen an expression library of C. trachomatis serovar L2 DNA. Four clones were recognized only by antisera from infected monkeys. Sequence analysis revealed that three of these immunoreactive clones overlap a common ORF, designated ORF D242 (encoding p242), in the C. trachomatis genome database. The fourth clone contains two complete ORFs, each encoding 32 kDa proteins that share identity with Treponema pallidum TroA and TroB (ORFs D067 and D068 in the C. trachomatis database, respectively). Immunoblot analysis of Escherichia coli lysates expressing C. trachomatis TroA, TroB and p242 fusion proteins showed that p242 and TroA, but not TroB, were detected by the sera collected from infected primates. Antibodies directed at TroA and p242 were also detected in sera from several C. trachomatis-infected patients, demonstrating that these proteins are also recognized by humans following infection. Immunoblot analysis with antibody against TroA and p242 also demonstrated that both antigens are present in higher abundance in infected ChoK1 cells relative to purified C. trachomatis EBs. Immunofluorescence microscopy shows that TroA and p242 are both localized to intracellular developmental forms at the margins of growing inclusions. Collectively, these studies identify two C. trachomatis proteins that are under-represented in EBs and are recognized uniquely in the context of infection.


Culture-Free Detection of Antibiotic Resistance Markers from Native Patient Samples by Hybridization Capture Sequencing.
The increasing incidence of antimicrobial resistance (AMR) is a major global challenge. Routine techniques for molecular AMR marker detection are largely based on low-plex PCR and detect dozens to hundreds of AMR markers. To allow for comprehensive and sensitive profiling of AMR markers, we developed a capture-based next generation sequencing (NGS) workflow featuring a novel AMR marker panel based on the curated AMR database ARESdb. Our primary objective was to compare the sensitivity of target enrichment-based AMR marker detection to metagenomics sequencing. Therefore, we determined the limit of detection (LOD) in synovial fluid and urine samples across four key pathogens. We further demonstrated proof-of-concept for AMR marker profiling from septic samples using a selection of urine samples with confirmed monoinfection. The results showed that the capture-based workflow is more sensitive and requires lower sequencing depth compared with metagenomics sequencing, allowing for comprehensive AMR marker detection with an LOD of 1000 CFU/mL. Combining the ARESdb AMR panel with 16S rRNA gene sequencing allowed for the culture-free detection of bacterial taxa and AMR markers directly from septic patient samples at an average sensitivity of 99%. Summarizing, the newly developed ARESdb AMR panel may serve as a valuable tool for comprehensive and sensitive AMR marker detection.


Biochemical response to ursodeoxycholic acid predicts survival in a North American cohort of primary biliary cirrhosis patients.
Biochemical response to ursodeoxycholic acid among patients with primary biliary cirrhosis remains variable, and there is no agreement of an ideal model. Novel assessment of response coupled to histologic progression was recently defined by the Toronto criteria. We retrospectively assessed transplant-free survival and clinical outcomes associated with ursodeoxycholic acid response to evaluate the Toronto criteria using a large North American cohort of PBC patients. Three hundred and ninety-eight PBC patients from the Mayo Clinic PBC Genetic Epidemiology Registry were assessed for ursodeoxycholic acid treatment and biochemical response per the Toronto criteria. Responders were defined by reduction in alkaline phosphatase to less than or equal to 1.67 times the upper normal limit by 2 years of treatment, whereas non-responders had alkaline phosphatase values greater than 1.67 times the upper normal limit. Probability of survival was estimated using the Kaplan-Meier method. Three hundred and two (76 %) patients were responders and 96 (24 %) were non-responders. Significantly more non-responders developed adverse events related to chronic liver disease compared to responders (hazard ratio (HR) 2.77, P = 0.001). Biochemical responders and early-stage disease at treatment start was associated with improved overall transplant-free survival compared to non-responders (HR 1.9) and patients with late-stage disease (HR 2.7) after age and sex adjustment. The Toronto criteria are capable of identifying ursodeoxycholic acid-treated primary biliary cirrhosis patients at risk of poor transplant-free survival and adverse clinical outcomes. Our data reveal that despite advanced disease at diagnosis, biochemical response per the Toronto criteria associates with improved overall transplant-free survival.


Hemorrhagic pituitary adenomas: clinicopathological features and surgical treatment.
Forty-five (9.9%) of 453 pituitary adenomas operated on between January 1973 and November 1988 demonstrated hemorrhagic changes at surgery: 24 had a blood collection, 12 had a blood collection associated with hemorrhagic necrosis, and 9 had hemorrhagic necrosis. Thirteen patients (28.9%) experienced the acute symptoms of pituitary apoplexy, whereas another 32 had an "asymptomatic" hemorrhage, that is, the clinical course was comparable to an uncomplicated adenoma. Nineteen tumors (42.2%) showed marked suprasellar extension, 8 (17.8%) showed moderate extension, and 11 (24.5%) showed slight extension; another 2 (4.4%) were laterosellar and 5 (11.1%) were intrasellar. Invasive behavior was present in 32 cases (71.1%) and this may suggest another hypothesis to explain the pathogenesis of tumoral hemorrhage. The incidence of hemorrhagic complications in invasive adenomas with marked suprasellar extension was particularly impressive; therefore, we do not suggest preoperative bromocriptine treatment in this type of tumor. Two of 14 patients operated on by the transcranial route died after surgery, whereas there was no operative mortality in the 31 patients operated on by the transsphenoidal route. It proved advantageous to operate as early as possible, even during the acute phase of pituitary apoplexy. The transsphenoidal approach gave the best results, but to achieve satisfactory late results multidisciplinary treatment was necessary, namely, postoperative radiotherapy in 23 patients, bromocriptine in 12, and endocrine replacement therapy in almost all. In an average follow-up period of 6.2 years, 5 (11.1%) symptomatic recurrences were observed.


Defibrotide for the treatment of severe hepatic veno-occlusive disease and multiorgan failure after stem cell transplantation: a multicenter, randomized, dose-finding trial.
Therapeutic options for severe hepatic veno-occlusive disease (VOD) are limited and outcomes are dismal, but early phase I/II studies have suggested promising activity and acceptable toxicity using the novel polydisperse oligonucleotide defibrotide. This randomized phase II dose-finding trial determined the efficacy of defibrotide in patients with severe VOD following hematopoietic stem cell transplantation (HSCT) and identified an appropriate dose for future trials. Adult and pediatric patients received either lower-dose (arm A: 25 mg/kg/day; n = 75) or higher-dose (arm B: 40 mg/kg/day; n = 74) i.v. defibrotide administered in divided doses every 6 hours for > or =14 days or until complete response, VOD progression, or any unacceptable toxicity occurred. Overall complete response and day +100 post-HSCT survival rates were 46% and 42%, respectively, with no significant difference between treatment arms. The incidence of treatment-related adverse events was low (8% overall; 7% in arm A, 10% in arm B); there was no significant difference in the overall rate of adverse events between treatment arms. Early stabilization or decreased bilirubin was associated with better response and day +100 survival, and decreased plasminogen activator inhibitor type 1 (PAI-1) during treatment was associated with better outcome; changes were similar in both treatment arms. Defibrotide 25 or 40 mg/kg/day also appears effective in treating severe VOD following HSCT. In the absence of any differences in activity, toxicity or changes in PAI-1 level, defibrotide 25 mg/kg/day was selected for ongoing phase III trials in VOD.


Connexins regulate calcium signaling by controlling ATP release.
Forced expression of gap junction proteins, connexins, enables gap junction-deficient cell lines to propagate intercellular calcium waves. Here, we show that ATP secretion from the poorly coupled cell lines, C6 glioma, HeLa, and U373 glioblastoma, is potentiated 5- to 15-fold by connexin expression. ATP release required purinergic receptor-activated intracellular Ca2+ mobilization and was inhibited by Cl- channel blockers. Calcium wave propagation also was reduced by purinergic receptor antagonists and by Cl- channel blockers but insensitive to gap junction inhibitors. These observations suggest that cell-to-cell signaling associated with connexin expression results from enhanced ATP release and not, as previously believed, from an increase in intercellular coupling.


Rapid sampling and long-term storage of subcutaneous adipose-tissue biopsies for determination of fatty acid composition.
The fatty acid composition of fat tissue is a valid indicator of the fatty acid composition of the diet over the preceding 1 to 3 yr. Here we describe a rapid method for sampling of buttock fat without anesthesia. On average, 35 mg can be obtained using a common blood-sampling system. Sampling of 40 subjects takes about 3 h. The procedure caused no more anxiety or discomfort than a routine venipuncture. Application in about 500 subjects has as yet yielded no infectious or other complications. With regard to the fatty acid composition of the biopsy, sampling site (left versus right buttock) was not found to be a source of error. The biopsies could be stored for periods up to 1.5 yr without changes in the fatty acid profile. With this method one can obtain biopsies from a large number of subjects and determine objectively the long-term fatty acid composition of their diet.


Clinical factors for choosing active surveillance: an analysis of papillary thyroid microcarcinoma patients with recurrence.
Active surveillance (AS) has been suggested as a management option for low-risk papillary thyroid microcarcinoma (PTMC). However, the currently proposed selection criteria for AS application do not consider various clinical factors. The purpose of this study was to analyze clinical factors related to recurrence that could be confirmed preoperatively in patients who underwent surgery for PTMC and to identify factors worth considering when deciding whether to apply AS. Data were collected from patients with PTMC who underwent surgical treatment at Chungnam National University Hospital. A retrospective cohort was established according to the presence or absence of recurrence during the follow-up period. In total, 2,717 patients were enrolled, of whom 60 experienced recurrence. Various clinical factors that could be identified before surgery were analyzed. The relationship between various clinical factors that could be confirmed preoperatively and recurrence was confirmed through Cox regression analysis and Kaplan-Meier curve analysis. BRAF mutation and the tall cell variant were significantly more common in patients with recurrence. In patients aged 55 years or older, the risk of recurrence was lower than in younger patients, while the recurrence-free survival (RFS) rate was higher. When choosing between surgical treatment or AS in PTMC patients, additional consideration of the patient's clinical factors, such as age and BRAF mutation status, may be required in addition to the existing criteria.


Status epilepticus in adults.
Status epilepticus is a common neurological emergency with considerable associated health-care costs, morbidity, and mortality. The definition of status epilepticus as a prolonged seizure or a series of seizures with incomplete return to baseline is under reconsideration in an effort to establish a more practical definition to guide management. Clinical research has focused on early seizure termination in the prehospital setting. The approach of early escalation to anaesthetic agents for refractory generalised convulsive status epilepticus, rather than additional trials of second-line anti-epileptic drugs, to avoid neuronal injury and pharmaco-resistance associated with prolonged seizures is gaining momentum. Status epilepticus is also increasingly identified in the inpatient setting as the use of extended electroencephalography monitoring becomes more commonplace. Substantial further research to enable early identification of status epilepticus and efficacy of anti-epileptic drugs will be important to improve outcomes.


[Global lessons of the Mexican health reform: empowerment through the use of evidence].
This paper illustrates, using as an example the recent reform of the Mexican health system, the potential of knowledge in the design and implementation of public policies. In the first part the relationship between knowledge and health is described. In part two, the efforts in Mexico to generate evidence that would eventually nourish the design and implementation of health policies are discussed. In the following sections the content and the guiding concept of the reform, the democratization of health, are analyzed. The paper concludes with the discussion of the main global lessons of this reform experience.


Perception of the ethical acceptability of live prey feeding to aquatic species kept in captivity.
Previous research into public perceptions of live prey feeding has been focused on terrestrial animals. The reasons for this likely relate to the difficulty humans have in being compassionate to animals who are phylogenetically distantly related. In order to test these assumptions, the general public (two groups; one who had just visited an aquarium; and one group who had just visited a zoo), aquarium professionals in the UK/US and terrestrial zoo animal professionals (UK) were investigated to see how they would differ in their responses when asked about feeding various live aquatic animals to one another. Likert based surveys were used to obtain data face to face and via online social media. Demographics in previous research identified a lower acceptance of live prey feeding by females, however in aquatic animals this was not reflected. Instead, separations in perception were seen to exist between participants dependent on whether they had just visited a zoo or aquarium, or worked with animals.


Placing a Central Vascular Access Device in a Patient With Substance Use Disorder: The Ethical Position of the Infusion Nurse.
When infusion nurses place central vascular access devices in patients with substance use disorder (SUD), they are both enabling treatment and making the patient more vulnerable to his or her addictive illness. Using the lens of rescue enables an exploration of the ethical position of the infusion nurse regarding these patients, even though rescue, per se, is inadequate to the complexity of the situation. Suggestions are offered to both the infusion nurse and the health care team for improving their ethical stance, as well as their care of patients with SUD.


Defendants with repeated competency evaluations.
In order to clarify the nature of those defendants referred multiple times for competency-to-stand-trial evaluations, 390 consecutive admissions to the Yale-New Haven Psychiatric Court Clinic were reviewed. Fifty-four of these had had more than one evaluation in the 2.5-year period under study and will be referred to as repeaters. Repeaters were significantly different from nonrepeaters in terms of diagnosis (p less than .01), tending to be more likely to be schizophrenic or affectively disordered. Repeaters also tended to have less education (p less than .01) and to be found competent less often (p less than .01) than nonrepeaters. No repeater-nonrepeater differences were found in age, race, or legal charge. Implications for future research on and management of this population are discussed.


Complete metabolic response is not uniformly predictive of complete pathologic response after induction therapy for esophageal cancer.
Positron emission tomography scanning is used to assess response to induction therapy after treatment of esophageal cancer. A decrease in standardized uptake value has been correlated with response to therapy, with a standardized uptake value of zero often assumed to indicate complete absence of disease. We hypothesize that a significant number of patients may have residual esophageal cancer despite complete metabolic response (CMR). A prospective database was reviewed for esophageal cancer patients receiving induction therapy followed by esophagectomy on whom both preinduction and postinduction positron emission tomography scans were obtained. Patients with a postinduction SUV of 0 (or "no uptake") were categorized as complete metabolic responders. Survival was calculated by the Kaplan-Meier statistic. Among 120 patients, 32 (27%) had postinduction CMR after chemotherapy (21 of 81, 26%) or chemoradiation (11 of 39, 28%). At surgery, 19 patients (59%) with CMR had residual disease, including 12 (38%) with nodal metastases. Even among patients with a negative postinduction biopsy, 4 of 10 (40%) had residual disease. Final pathologic stages of patients with CMR were yp0 (complete pathologic response) in 13 (41%), ypI in 4 (12%), ypII in 9 (28%), and ypIII in 6 (19%). Three-year survival was 83% in the CMR group versus 41% in the remainder of the cohort (p = 0.02). A CMR on postinduction positron emission tomography scan predicts but should not be assumed to be synonymous with complete pathologic response in esophageal cancer patients. The presence of residual disease should be strongly considered despite CMR and negative biopsy in patients receiving induction therapy.


Pediatric mortality in Africa: plasmodium falciparum malaria as a cause or risk?
The disability adjusted life year (DALY) approach of defining cause-specific health burdens is becoming the benchmark for international disease control prioritization. For malaria, this categorical approach may not fully capture its burden that includes chronic anemia, low birth weight, and enhancement of the severity of other childhood diseases. We investigated the extent to which malaria acts as a risk factor for all-cause mortality in African children less than five years of age from 1) ecologic associations between Plasmodium falciparum infection prevalence (PR) and under-five mortality, and 2) reductions in all-cause under-five mortality achieved in malaria intervention trials. Across 48 demographic surveillance studies, when adjusted for secular trends, PR more than doubled all-cause mortality (P = 0.0001). Trials of insecticide-treated mosquito nets generally found smaller population-attributable fractions of pediatric mortality to malaria infection, which may relate to their imperfect coverage and efficacy. In conclusion, the disability and death burden due to malaria in African children could be higher than that detectable from cause-specific DALY estimations.


Helical conformations of hexapeptides containing N-terminus diproline segments.
The role of N-terminus diproline segments in facilitating helical folding in short peptides has been investigated in a set of model hexapeptides of the type Piv-Xxx-Yyy-Aib-Leu-Aib-Phe-OMe (Piv, pivaloyl). Nine sequences have been investigated with the following N-terminus dipeptide segments: (D)Pro-Ala (4) and Pro-PsiPro (5, Psi, pseudoproline), Ala-Ala (6), Ala-Pro (7), Pro-Ala (8), Aib-Ala (9), Ala-Aib (10). The analog sequences Piv-Pro-Pro-Ala-Leu-Aib-Phe-OMe (2) and Piv-Pro-Pro-Ala-Aib-Ala-Aib-OMe (3) have also been studied. Solid state conformations have been determined by X-ray crystallography for peptides 4, 6, and 8 and compared with the previously determined crystal structure of peptide 1 (Boc-Pro-Pro-Aib-Leu-Aib-Val-OMe); (Rai et al., JACS 2006, 128, 7916-7928). Peptides 1 and 6 adopt almost identical helical conformations with unfolding of the helix at the N-terminus Pro (1) residue. Peptide 4 reveals the anticipated (D)Pro-Ala type II' beta-turn, followed by a stretch of 3(10)-helix. Peptide 8 adopts a folded conformation stabilized by four successive 4-->1 intramolecular hydrogen bonds. Ala (2) adopts an alpha(L) conformation, resulting in a type II beta-turn conformation followed by a stretch of 3(10)-helix. Conformational properties in solution were probed using solvent perturbation of NH chemical shifts which permit delineation of hydrogen bonded NH groups and nuclear Overhauser effects (NOEs) between backbone protons, which are diagnostic of local residue conformations. The results suggest that, continuous helical conformations are indeed significantly populated for peptides 2 and 3. Comparison of the results for peptides 1 and 2, suggest that there is a significant influence of the residue that follows diproline segments in influencing backbone folding.


Development and Validation of an Atrial Fibrillation Knowledge Assessment Tool (AFKAT).
Assessing and improving public knowledge of atrial fibrillation (AF) could increase its detection rate and the subsequent use of stroke prevention therapies. However, there is no validated AF knowledge assessment tool applicable to the general population, including those at risk of AF. Therefore, we aimed to develop and validate such a tool. The tool was developed from a literature review and discussion with subject matter experts. Content validity was ensured by a ten-member panel of experts comprising cardiologists and pharmacists. An online validation survey was conducted and reported based on the Checklist for Reporting Results of Internet E-Surveys (CHERRIES). The survey evaluated the tool performance by construct validity, internal consistency reliability, item discrimination, difficulty index and ease of readability. The survey participants included 14 general medical specialists, 20 fourth-year and 33 second-year undergraduate pharmacy students, and 122 members of the general public. The tool had satisfactory content validity, with a scale content validity index of 0.8. The mean percentage knowledge scores for general medical specialists and fourth-year pharmacy students were higher than second-year pharmacy students, followed by the general public (92.9%, 87.6%, 68.5% and 53.4%, respectively; p-value < 0.001), supporting construct validity. The tool had good internal consistency reliability (Cronbach's alpha = 0.91). The item-total correlation was in the preferred range of 0.23 to 0.71. The Atrial Fibrillation Knowledge Assessment Tool is a valid instrument and can be used to investigate AF knowledge of the general population.


Immunohistological studies on the distribution of cytochrome b3 reductase in rat liver microsomes.
The distribution of cytochrome b5 reductase in rat liver microsomes purified by density gradient centrifugation was studied using anti-cytochrome b5 reductase/anti-ferritin hybrid antibodies labeled with ferritin. Electron micrographs show that the outer surface and the polar cap of tangential sections of essentially all vesicles derived from endoplasmic reticulum are specifically marked with a few ferritin grains which are not localized in large patches but arranged randomly. This finding was correlated with morphometrical and biochemical measurements. The results, supporting our earlier investigations, suggest that cytochrome b5 reductase may be clustered at the most in very small assemblies consisting of a few molecules which, in turn, are statistically uniformly distributed over the total membrane system.


Quantification of adducts formed in DNA treated with N-acetoxy-2-acetylaminofluorene or N-hydroxy-2-aminofluorene: comparison of trifluoroacetic acid and enzymatic degradation.
We have examined two methods of preparation of DNA adducts from phi X174 RF DNA modified by [3H]N-acetoxy-2-acetylaminofluorene ([3H]NA-AAF) or N-hydroxy-2-aminofluorene ([3H]N-OH-AF). Hydrolysis by enzymes (DNase I, snake venom phosphodiesterase and alkaline or acid phosphatase) and subsequent reverse phase h.p.l.c. of phi X174 RF DNA treated with [3H]NA-AAF yielded 73% N-(deoxyguanosin-8-yl)-2-acetylaminofluorene (dG-C8-AAF), 7% 3-(deoxyguanosin-N2-yl)-2-acetylaminofluorene (dG-N2-AAF), and a peak of unidentified radioactivity (13%). When [3H]N-OH-AF modified phi X174 DNA was analyzed, both N-(deoxyguanosin-8-yl)-2-aminofluorene (dG-C8-AF) and a large percentage of the imidazole ring-opened derivative and unidentified products were found. In contrast, when anhydrous trifluoroacetic acid (TFA) was used to degrade these DNAs, we found for the [3H]NA-AAF modified DNA 86% N-(guanin-8-yl)-2-acetylaminofluorene (G-C8-AAF) and 6% 3-(guanin-N2-yl)-2-acetylaminofluorene (G-N2-AAF), while for [3H]N-OH-AF modified DNA only the N-(guanin-8-yl)-2-aminofluorene (G-C8-AF) was found. When DNA was prepared from human fibroblasts treated with [3H]NA-AAF, only the G-C8-AF product was obtained. Thus, anhydrous TFA solvolysis followed by reverse phase h.p.l.c. is a rapid and convenient method to obtain quantitative yields of DNA adducts formed with acetylaminofluorene and related compounds: quantification by this method prevents loss of G-N2-AAF adducts, the conversion of AAF adducts to AF adducts, and the production of ring opened products in guanine residue.


Psychological impact of the tsunami on children and adolescents from the andaman and nicobar islands.
The aim of this article is to present the assessment of the presentation of symptoms and psychiatric morbidity of children and adolescents from the Andaman and Nicobar islands during the first 3 months following the December 2004 earthquake and tsunami. According to predefined criteria, a primary survivor is one who was exposed directly to the earthquake and tsunami, a secondary survivor is one with close family and personal ties to primary survivors, and tertiary survivors are individuals from the communities beyond the impact area, a majority of which were exposed to the earthquake only. This study included 37 primary and secondary survivors (aged </= 18 years) and 498 tertiary survivors of the tsunami disaster. Tertiary survivors were recruited from the 10th and 12th grades of schools in Port Blair, India. The following 3 screening and treatment methods were adopted: (1) mental health clinic, (2) art therapy, and (3) group discussions. The most common psychiatric morbidities observed among the primary and secondary survivors were adjustment disorder (N = 5, 13.5%), depression (N = 5, 13.5%), panic disorder (N = 4, 10.8%), posttraumatic stress disorder (N = 4, 10.8%), schizophrenia (N = 1, 2.7), and other disorders (N = 16, 43.2%). Subclinical syndrome was present in the majority of the primary and secondary survivors. Few tertiary survivors had subsyndromal symptoms. Only a few of the primary and secondary survivors required intensive individual psychiatric interventions; however, a majority of the primary, secondary, and tertiary survivors required community-based group interventions. Community-based group interventions and group discussions are simple, easy to implement using local resources, and effective in all groups, and provide important components of psychosocial rehabilitation. This kind of approach should be started as early as possible, targeting all children and adolescents affected by any disaster in developing countries.


CRISPR based editing of SIV proviral DNA in ART treated non-human primates.
Elimination of HIV DNA from infected individuals remains a challenge in medicine. Here, we demonstrate that intravenous inoculation of SIV-infected macaques, a well-accepted non-human primate model of HIV infection, with adeno-associated virus 9 (AAV9)-CRISPR/Cas9 gene editing construct designed for eliminating proviral SIV DNA, leads to broad distribution of editing molecules and precise cleavage and removal of fragments of the integrated proviral DNA from the genome of infected blood cells and tissues known to be viral reservoirs including lymph nodes, spleen, bone marrow, and brain among others. Accordingly, AAV9-CRISPR treatment results in a reduction in the percent of proviral DNA in blood and tissues. These proof-of-concept observations offer a promising step toward the elimination of HIV reservoirs in the clinic.


Early-life stress, depressive symptoms, and inflammation: the role of social factors.
To identify modifiable, social factors that moderate the relationship between early-life stress (ELS) and health outcomes as measured by depressive symptoms and inflammation. Data were from 3,416 adults (58.28% female), ages 36 - 97 (Mage = 68.41; SDage = 10.24) who participated in the 2006 wave of the Health and Retirement Study, a nationally representative sample of older adults in the United States. This study used hierarchical regression analyses to first test the main effects of ELS on depressive symptoms and inflammation (high-sensitivity C-reactive protein). Four social factors (perceived support, frequency of social contact, network size, and volunteer activity) were assessed as moderators of the ELS-depression and ELS-inflammation relationships. There was a small, positive association between ELS and depressive symptoms (B = 0.17, SE = 0.05, p = .002), which was moderated by social contact and perceived support. Specifically, ELS was only associated with elevated depressive symptoms for participants with limited social contact (B = 0.24, SE = 0.07, p < .001) and low perceived support (B = 0.24, SE = 0.07, p < .001). These associations remained after accounting for potential confounds (age, body-mass index, adulthood stress, and marital status). Increased social contact and perceived support may be protective for individuals at a higher risk of developing depressive symptoms as a result of ELS. Future interventions may benefit from leveraging these social factors to improve quality of life in adults with ELS.


Binding of insulin to bovine liver plasma membrane. Use of insulin analogues modified at the A1 residue.
Bovine liver plasma membranes [Rösen, Ehrich, Junger, Bubenzer & Kühn (1979) Biochim. Biophys. Acta587, 593-605] show similar insulin-binding characteristics, as evaluated by Scatchard analysis, to those of membrane systems from other species. However, the dissociation rate of bound insulin cannot be accelerated by the addition of insulin, in contrast with membranes isolated from rat liver. The dissociation rate is strongly dependent on the pH. Although dependent on temperature, the total capacity of binding sites is minimally changed, but the number of high-affinity sites is increased 2-3-fold, by lowering the incubation temperature. These data might be interpreted by assuming a single population of receptors whose distribution between different affinity states depends on temperature. In competition studies, most of the modified insulins examined show a close correlation between binding, determined in plasma membranes from bovine liver, and biological activity, measured in adipocytes. The hypothesis that a positive charge on the A1 residue may be favourable for binding is supported by experiments with an isosteric pair of insulins modified at this residue ([carbamoyl-Gly(A1)]- and [amidino-Gly(A1)]insulin) and with modified insulins carrying one or more positive charges on the A1 residue ([Arg-Gly(A1)]-, [Arg-Arg-Gly(A1)]-, [Arg-Arg-Arg-Gly(A1)]- and [Lys-Arg-Gly(A1)]insulin). The latter insulin derivatives show a higher binding activity for plasma membranes from bovine, porcine and rat liver than expected from their biological activities in adipocytes.


Differentiating factors between erythropoiesis-stimulating agents: an update to selection for anaemia of chronic kidney disease.
Erythropoiesis-stimulating agents (ESAs) have become a hallmark of anaemia therapy in patients with chronic kidney disease (CKD). Although different ESAs are available for the treatment of renal anaemia, each nephrologist should select a single ESA for an individual patient. Epoetin alfa and epoetin beta have been used 1-3 times weekly but extended-interval dosing up to every 4 weeks is also effective in a substantial majority of CKD patients. However, the epoetin dose necessary to achieve or maintain target haemoglobin (Hb) levels increases substantially as the dosing interval increases. Subcutaneous administration of short-acting ESAs is more effective than the intravenous route of administration. Darbepoetin alfa and the continuous erythropoietin receptor activator (CERA) have been developed as a treatment for anaemia with extended administration intervals (every 2 weeks and every 4 weeks, respectively). Dose requirements for these long-acting ESAs are independent of the route of administration. Patents of short-acting ESAs have expired, which has opened the field for biosimilars. Epoetin biosimilars approved by the European Medicines Agency (EMA) or the US Food and Drug Administration (FDA) have been shown to have a comparable efficacy and safety profile to their originators. An alarming increase in pure red cell aplasia (PRCA) in Thailand with follow-on epoetins manufactured in Asia (but also those manufactured in Latin America) indicates that stringent country-specific approval and pharmacovigilance protocols for ESAs manufactured in non-North American and non-EU European countries are urgently needed. Two PRCA cases occurring with subcutaneous HX575 (one certain, one likely) indicate that chances of inducing a more immunogenic product are unpredictable, even with a biosimilar epoetin approved under the EMA biosimilar approval pathway. Phase III clinical trials with peginesatide, a pegylated synthetic peptide-based ESA without any homology to erythropoietin raised safety concerns in non-dialysis CKD patients but not in dialysis patients.


Identification of conductive hearing loss using air conduction tests alone: reliability and validity of an automatic test battery.
The primary objective of this study was to determine whether a combination of automatically administered pure-tone audiometry and a tone-in-noise detection task, both delivered via an air conduction (AC) pathway, could reliably and validly predict the presence of a conductive component to the hearing loss. The authors hypothesized that performance on the battery of tests would vary according to hearing loss type. A secondary objective was to evaluate the reliability and validity of a novel automatic audiometry algorithm to assess its suitability for inclusion in the test battery. Participants underwent a series of hearing assessments that were conducted in a randomized order: manual pure-tone air conduction audiometry and bone conduction audiometry; automatic pure-tone air conduction audiometry; and an automatic tone-in-noise detection task. The automatic tests were each administered twice. The ability of the automatic test battery to: (a) predict the presence of an air-bone gap (ABG); and (b) accurately measure AC hearing thresholds was assessed against the results of manual audiometry. Test-retest conditions were compared to determine the reliability of each component of the automatic test battery. Data were collected on 120 ears from normal-hearing and conductive, sensorineural, and mixed hearing-loss subgroups. Performance differences between different types of hearing loss were observed. Ears with a conductive component (conductive and mixed ears) tended to have normal signal to noise ratios (SNR) despite impaired thresholds in quiet, while ears without a conductive component (normal and sensorineural ears) demonstrated, on average, an increasing relationship between their thresholds in quiet and their achieved SNR. Using the relationship between these two measures among ears with no conductive component as a benchmark, the likelihood that an ear has a conductive component can be estimated based on the deviation from this benchmark. The sensitivity and specificity of the test battery vary depending on the size of this deviation, but increase with increasing ABG size, with decreasing test frequency, and when results from multiple test frequencies are taken into account. The individual automatic tests comprising the battery were found to be reliable and valid, with strong, significant correlations between the test and retest results (r = 0.81 to 0.99; p < 0.0001) and between automatic and manual audiometry procedures (r = 0.98 to 0.99; p < 0.0001). The presence of an ABG can be predicted with a reasonably high degree of accuracy using AC tests alone. Applications of such a test battery include any clinical context in which bone conduction audiometry or specialized diagnostic equipment is unavailable or impractical. Examples of these include self-fitting hearing aids, whose efficacy relies on the ability of the device to automatically administer an in situ hearing test; self-administered adult hearing screenings in both clinical and home environments; large-scale industrial hearing conservation programs; and test environments in which ambient noise levels exceed the maximum permissible levels for unoccluded ears.


[Proton pump inhibitors when using clopidogrel: balance between necessity and disadvantages].
Rationale When patients are using carbasalate calcium or acetylsalicylic acid (ASA), it is recommended to prescribe a proton pump inhibitor (PPI) in order to prevent gastrointestinal (GI) bleeding. Should this recommendation also be followed for patients who are using clopidogrel in monotherapy, which is increasingly the case in practice? Method In a systematic literature review of the occurrence of GI bleeding when using clopidogrel versus ASA, we included 9 studies that compared the risk of GI bleeding when using ASA with clopidogrel monotherapy. Results These 9 studies on clopidogrel and ASA show that the risk of GI bleeding is also elevated when using clopidogrel monotherapy and that it is comparable with the risk of GI bleeding when using ASA. Conclusion Based on the current literature, we recommend prescribing pantoprazole to patients who are using clopidogrel monotherapy and have additional risk factors for GI bleeding, in accordance with the procedure for low-dose ASA. The risk of GI bleeding must be weighed against the disadvantages of using PPIs.


Anticonvulsant drug therapy after aneurysmal subarachnoid hemorrhage: a critically appraised topic.
Seizures are a complication of aneurysmal subarachnoid hemorrhage (aSAH). To evaluate whether antiepileptic drug (AED) prophylaxis after aSAH reduces seizure risk and whether it is associated with improved neurological outcomes. The objective was addressed through the development of a critically appraised topic that included a clinical scenario, structured question, search strategy, critical appraisal, assessment of results, evidence summary, commentary, and bottom line conclusions. Neurology consultants and residents, a medical librarian, clinical epidemiologists, and content experts in the fields of epilepsy, neurosurgery, and critical care contributed to the review and placed the evidence in clinical context. There were no relevant randomized, controlled trials that addressed the question. A post hoc analysis of data from 4 trials of tirilazad for aSAH showed that prophylactic AED therapy was associated with worse Glasgow Outcome Scale scores at 3 months (odds ratio 1.56, 95% confidence interval 1.16-2.10; P = 0.003) but numerous confounders limit data interpretation. There are insufficient data to support or refute the prophylactic use of AED therapy after aSAH. Randomized, controlled trials are needed to address the efficacy and risks of AEDs in this setting and should take into account factors such as aneurysmal factors (location, hemorrhage grade, degree of parenchymal injury), type of aneurysm surgery (clip vs. coil), and evaluate the timing and duration of AED use.


Outcomes of a Tailored Intervention for Cigarette Smoking Cessation Among Latinos Living With HIV/AIDS.
Tobacco use has emerged as a leading killer among persons living with HIV, with effective approaches to tobacco treatment still unknown. HIV infection is nearly 3 times as prevalent in Latinos than in non-Latino Whites. This study reports the results of a randomized trial comparing a tailored intervention to brief counseling for smoking cessation among Latino smokers living with HIV (LSLWH). LSLWH (N = 302; 36% female, 10% employed full-time, 49% born in United States) were randomized to 4 in-person sessions of a tailored intervention (Aurora) or 2 in-person sessions of brief advice (enhanced standard care [ESC]). Both groups received 8 weeks of nicotine replacement therapy (NRT) patch. Biochemically validated 6- and 12-month 7-day point-prevalence abstinence (PPA) rates were compared, along with secondary outcomes (e.g., reduction to light smoking, NRT adherence). Seven-day PPA rates reached 8% versus 11% at 6 months and 6% versus 7% at 12 months, for Aurora and ESC, respectively, with no between-group differences (p values > .40). Significant changes from baseline to 6 and 12 months among intervention targets were noted (percentage reduction in heavy smoking and dependence; increases in knowledge and self-efficacy). Baseline smoking frequency, older age, and higher intensity of patch use during the trial emerged as significant predictors of abstinence at 6 months. There was no evidence that the tailored intervention improved cessation rates. Interventions that encourage use of, and adherence to, empirically validated cessation aids require further development to reduce tobacco-related death and disease in this vulnerable population.


Serotonin's role in alcohol's effects on the brain.
Serotonin is an important brain chemical that acts as a neurotransmitter to communicate information among nerve cells. Serotonin's actions have been linked to alcohol's effects on the brain and to alcohol abuse. Alcoholics and experimental animals that consume large quantities of alcohol show evidence of differences in brain serotonin levels compared with nonalcoholics. Both short- and long-term alcohol exposure also affect the serotonin receptors that convert the chemical signal produced by serotonin into functional changes in the signal-receiving cell. Drugs that act on these receptors alter alcohol consumption in both humans and animals. Serotonin, along with other neurotransmitters, also may contribute to alcohol's intoxicating and re. warding effects, and abnormalities in the brain's serotonin system appear to play an important role in the brain processes underlying alcohol abuse.


Pressor responses to vasopressin in rabbits with 3-day renal artery stenosis.
One-kidney rabbits were subjected to renal artery stenosis, and acute experiments were performed 3 days later on conscious animals; one-kidney rabbits without renal artery stenosis served as controls. Rabbits with 3-day renal artery stenosis were normotensive and had normal values for plasma renin activity. Intravenous infusion of arginine vasopressin at 5 mU.min-1.kg body wt-1 for 5 min resulted in a significantly (P less than 0.01) greater increase in mean arterial pressure and total peripheral resistance (TPR) in the renal artery stenosis rabbits than in the controls. Infusion of the angiotensin II (AII) competitive antagonist, [Sar1, Ile8]AII, before the vasopressin infusion abolished the hyperresponsiveness to vasopressin in the renal artery stenosis rabbits and resulted in changes in mean arterial pressure and TPR that were approximately of the same magnitude as the controls. Infusion of [SAr1, Ile8]AII before vasopressin infusion in control rabbits did not alter the cardiovascular responses to vasopressin. Because previous studies have shown that 3-day renal artery stenosis rabbits have exaggerated pressor responses to norepinephrine and that this hyperresponsiveness to norepinephrine is blocked by [Sar1, Ile8]-AII, the present study with vasopressin provided evidence that the increased responsiveness in this model is not specific for a single pressor agent. These studies also demonstrated that AII plays an important role in mediating the exaggerated pressor responses to vasopressin in this prehypertensive model.


Mucous membrane pemphigoid with generalized blisters: IgA and IgG autoantibodies target both laminin-332 and type XVII collagen.
Mucous membrane pemphigoid (MMP) is a mucous membrane-dominated, subepidermal autoimmune blistering disease in which autoantibodies usually react with the C-terminal domain of type XVII collagen (COL17) or with laminin-332. Only a few cases of MMP with widespread blisters have been reported. Serologically, IgA and IgG class autoantibodies directed against COL17 or IgG autoantibodies directed against laminin-332 in patients with MMP have been well documented. MMP cases in which IgA reacts with laminin-332, however, are extremely rare. We report a case of MMP in a 67-year-old man. Clinical examination revealed extensive mucosal lesions as well as generalized blisters and erosions that healed with scar formation. The disease was intractable to treatment with systemic steroids. Interestingly, in addition to IgG directed against laminin-332 and the noncollagenous 16A (NC16A) and C-terminal domains of COL17, circulating IgA reacting with laminin-332 and with the NC16A domain of COL17 was also detected. This is the first MMP case with circulating IgA and IgG autoantibodies against both laminin-332 and COL17.


Liquid-crystal microlens arrays using patterned polymer networks.
A real-time dynamically tunable-focus microlens array made from a polymer-liquid-crystal (LC) composite is demonstrated. The polymer was first patterned in microlens array cavities by lamination, and the LC-monomer mixture was then injected to the molded polymer cavities and finally stabilized by UV light-induced networks. Using this new fabrication method, we demonstrated a lens with a spherical shape and a glazed surface. This LC-based microlens can reach approximately 100% light efficiency for linearly polarized light. The saturation voltage of the lens is approximately 60 Vrms, and the response time is approximately 30 ms.


Temperature-Dependent Lipid Extraction from Membranes by Boron Nitride Nanosheets.
Two-dimensional (2D) materials can mechanically insert into cell membranes and extract lipids out, thus leading to the destruction of cell integrity. On the one hand, the cytotoxicity of 2D materials can be harnessed in surface engineering to resist biofouling, and on the other hand, it causes great concern with in vivo biomedical applications ranging from drug delivery to nanoimaging. To understand the nature of this cytotoxic behavior and find strategies to control it, we performed molecular dynamics (MD) simulations on the lipid extraction of hexagonal boron nitride (BN) nanosheets from lipid membranes. Interestingly, we observed that the lipid extraction behavior suddenly disappears as temperature decreases. Structural analyses revealed that this temperature dependence is related to the lipid membrane phase transition, which was confirmed by an additional membrane model with phase state regulated by cholesterol. The potential of mean force calculation was adopted to clarify the thermodynamic origin of these results, which also indicates directions to adjust the lipid extraction behavior of nanomaterials. Overall, this work suggests that the cytotoxic mechanical interactions between 2D materials and cell membranes can be controlled by temperature and other factors which can induce phase transitions of lipid membranes and that the thermodynamic threshold of the lipid extraction varies for surfaces with different curvature. This work clarifies the thermodynamics in the lipid extraction phenomenon of 2D materials and indicates possible strategies to adjust the antibacterial performance or cytotoxicity of 2D materials.


Persistent chronic inflammation and infection by Chikungunya arthritogenic alphavirus in spite of a robust host immune response.
Alphaviruses, including Chikungunya virus (CHIKV), produce a transient illness in humans, but severe forms leading to chronic incapacitating arthralgia/arthritis have been reported by mechanisms largely ill-characterized. The pathogenesis of CHIKV was addressed in a prospective cohort study of 49 hospitalized patients from Reunion Island subsequently categorized into two distinct groups at 12 mo postinfection. Comprehensive analyses of the clinical and immunological parameters throughout the disease course were analyzed in either the "recovered" or the "chronic" groups to identify prognostic markers of arthritis-like pathology after CHIKV disease. We found that the chronic group consisted mainly of more elderly patients (>60 y) and with much higher viral loads (up to 10(10) viruses per milliliter of blood) during the acute phase. Remarkably, a rapid innate immune antiviral response was demonstrated by robust dendritic/NK/CD4/CD8 cell activation and accompanied by a rather weak Th1/Th2 cytokine response in both groups. Interestingly, the antiviral immune response witnessed by high levels of IFN-alpha mRNA in PBMCs and circulating IL-12 persisted for months only in the chronic group. CHIKV (RNA and proteins) was found in perivascular synovial macrophages in one chronic patient 18 mo postinfection surrounded by infiltrating NK and T cells (CD4(++) but rare cytotoxic CD8). Fibroblast hyperplasia, strong angiogenesis, tissue lesions given the high levels of matrix metalloproteinase 2, and acute cell death [high cleaved poly(ADP-ribose) polymerase staining] were observed in the injured synovial tissue. These observed cellular and molecular events may contribute to chronic arthralgia/arthritis targeted by methotrexate used empirically for effective treatment but with immunosuppressive function in a context of viral persistence.


Cultivation reveals physiological diversity among defensive 'Streptomyces philanthi' symbionts of beewolf digger wasps (Hymenoptera, Crabronidae).
'Candidatus Streptomyces philanthi' is a monophyletic clade of formerly uncultured bacterial symbionts in solitary digger wasps of the genera Philanthus, Philanthinus and Trachypus (Hymenoptera, Crabronidae). These bacteria grow in female-specific antennal reservoirs and - after transmission to the cocoon - produce antibiotics protecting the host larvae from fungal infection. However, the symbionts' refractoriness to cultivation has thus far hampered detailed in vitro studies on their physiology and on the evolutionary changes in metabolic versatility in response to the host environment. Here we isolated in axenic culture 22 'Streptomyces philanthi' biovars from different host species. Sequencing of gyrB revealed no heterogeneity among isolates within host individuals, suggesting low levels of (micro)diversity or even clonality of the symbionts in individual beewolf antennae. Surprisingly, however, isolates from different host species differed strongly in their physiology. All biovars from the Eurasian/African Philanthus and the South American Trachypus host species had high nutritional demands and were susceptible to most antibiotics tested, suggesting a tight association with the hosts. By contrast, biovars isolated from the genus Philanthinus and the monophyletic North American Philanthus clade were metabolically versatile and showed broad antibiotic resistance. Concordantly, recent horizontal symbiont transfer events - reflected in different symbiont strains infecting the same host species - have been described only among North American Philanthus species, altogether indicative of facultative symbionts potentially capable of a free-living lifestyle. Phylogenetic analyses reveal a strong correlation between symbiont metabolic versatility and host phylogeny, suggesting that the host environment differentially affects the symbionts' evolutionary fate. Although opportunistic bacteria were occasionally isolated from the antennae of different host species, only filamentous Actinobacteria (genera Streptomyces, Amycolatopsis and Nocardia) could replace 'S. philanthi' in the antennal gland reservoirs. Our results indicate that closely related bacteria from a monophyletic clade of symbionts can experience very different evolutionary trajectories in response to the symbiotic lifestyle, which is reflected in different degrees of metabolic versatility and host-dependency. We propose that the host-provided environment could be an important factor in shaping the degenerative metabolic evolution in the symbionts and deciding whether they evolve into obligate symbionts or remain facultative and capable of a host-independent lifestyle.